Electrochemical Nucleotide Detection via Intercalating Mediator

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Solution Overview

Problem

Existing electrochemical detection methods can only demonstrate and quantify the presence of target nucleotide sequences in biological samples, but not identify their nature effectively.

Innovation Solution

A method involving activatable amplification means with free nucleotides, specific primers, and an oxido-reducible compound that intercalates between replicated nucleotide sequences, causing a decrease in electric current, allowing for the identification and quantification of target nucleotide sequences through successive amplification cycles and thermal energy application.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If an oxidizing agent is used to react with oxidizable nucleotide bases during amplification, then the presence of target nucleic acid can be detected through current decrease, but the method cannot identify the nature of the target sequence

Engineering Contradiction:

Improvedetection capabilityVSAvoidsequence identification information

Core Design Contradiction:

Measurement precisionVSLoss of information

Solution Approach 1:

The patent introduces an oxido-reducible compound as an intermediary substance that specifically intercalates between nucleotide bases. This compound acts as a mediator that provides additional information about the target sequence nature while maintaining the detection function, thereby resolving the contradiction between detection capability and information loss.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent utilizes electrochemical signal changes (analogous to color changes in optical methods) of the oxido-reducible compound when it intercalates between nucleotide bases. The compound's electrochemical properties change based on its intercalation state, providing information about the target sequence nature through measurable signal variations.

Inventive Principle:

Principle #32Color changes

2Productivity

If nucleotides are consumed during amplification to produce replicated sequences, then target sequence presence can be quantified through current decrease, but the method lacks specificity for identifying different sequence types

Engineering Contradiction:

Improvequantification capabilityVSAvoidsequence identification specificity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The oxido-reducible compound serves as a specific intermediary that interacts differently with different nucleotide sequences. Its electrochemical response provides sequence-specific information that enhances both quantification accuracy and identification specificity, resolving the contradiction between productivity and reliability.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent exploits changes in electrochemical parameters of the oxido-reducible compound during intercalation. These parameter changes provide additional dimensions for distinguishing different sequence types, thereby improving both quantification and identification specificity simultaneously.

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

Enables the rapid detection and quantification of target nucleotide sequences while identifying their nature by measuring electric current changes during amplification cycles and thermal energy-induced release of the oxido-reducible compound, providing improved sensitivity and specificity compared to prior art methods.

Implementation Method 1

an oxido-reducible compound capable of intercalating, during the replication, between the nucleotides forming said replicated target sequences

Methodology Applied

Scientific EffectIntercalation:

Implementation Method 2

said replicated target sequences causing the inhibition of the electrochemical activity of said intercalated oxido-reducible compound

Methodology Applied

Scientific EffectElectrochemical activity inhibition:

Implementation Method 3

an electric field is applied to said sample in order to activate said oxido-reducible compound

Methodology Applied

Scientific EffectElectric field activation: Electric Field

Implementation Method 4

an oxido-reducible compound capable of reacting with respect to said nucleotides

Methodology Applied

Scientific EffectElectrochemical reaction: Redox Reactions

Implementation Method 5

said sample is supplied with thermal energy in such a way as to cause the release of said intercalated oxido-reducible compound

Methodology Applied

Scientific EffectThermal release: Heating

Data Source

PatentUS8962240B2Method for electrochemically identifying target nucleotide sequences

Publication Date: 2015.02.24 CENT NAT DE LA RECH SCI (C N R S)

AI summary

A method and assembly for electrochemically identifying target nucleotide sequences. The method includes supplying a biological sample that may contain a predetermined target nucleotide sequence; supplying activatable amplification materials comprising free nucleotides to form replicated target nucleotide sequences; supplying an oxido-reducible compound capable of being inserted during replication between the nucleotides forming the replicated target sequences; and activating the activatable amplification materials before applying an electric field to the sample in order to activate the oxido-reducible compound. The replicated target sequences cause inhibition of electrochemical activity of the inserted oxido-reducible compound, and the presence of the predetermined target nucleotide sequence is determined in instances where the electric current decreases.